Beware the Black Widow at Claim Time: A Report of Three Cases.

Moral hazard is well known to life insurance underwriters and medical directors to increase the risk of adverse consequences to insured individuals. The underwriting investigation of proposed insureds at time of policy issue is done to ensure no likely moral hazard exists. However, not all situations involving moral hazard may be identified at time of underwriting and policy issue, and may only be identified at time of claim. Three cases that were underwritten for life expectancies in legal matters are described here as examples of moral hazard identified at time of severe injury and/or death. All three of these cases involved a woman who manipulated her male partner into situations that increased the man's risk of severe injury and/or death to the woman's financial benefit. Such "black widows" made a great deal of effort over an extensive period of time to ensure that the moral hazard set up for their male partners resulted in a substantial financial windfall through litigation. The moral hazard set up by a black widow thus can be considered by the life insurance industry as sufficiently anti-selective and speculative to deny a claim at any time after policy issue.

Identification and Assessment of Undiagnosed Fetal Alcohol Spectrum Disorder: A Report of Three Cases.

Fetal alcohol spectrum disorder (FASD) and its associated physical and mental conditions is the most prevalent congenital impairment causing developmental and intellectual disability worldwide. Like alcohol abuse, FASD is typically undiagnosed by primary care providers. And like alcohol abuse, life underwriters and medical directors need to be aware of the signs, symptoms, and behaviors associated with FASD to accurately detect, identify, evaluate and assess the mortality risk. Three cases of suspected undiagnosed FASD that were underwritten for life expectancies in legal matters are discussed in this report. Not only were these patients' risks for excess mortality elevated due to their initial neurologic injury due to prenatal exposure to alcohol, but these cases demonstrate the importance of the stability and care needed to make them insurable. The following paper discusses the clinical and social settings at birth that may give underwriters and medical directors some clue to a potential case of the child having FASD and then to assess their statistical and lifestyle mortality risks.

Improving the stability of temporal statistics in transition path theory with sparse data.

Ulam's method is a popular discretization scheme for stochastic operators that involves the construction of a transition probability matrix controlling a Markov chain on a set of cells covering some domain. We consider an application to satellite-tracked undrogued surface-ocean drifting buoy trajectories obtained from the National Oceanic and Atmospheric Administration Global Drifter Program dataset. Motivated by the motion of Sargassum in the tropical Atlantic, we apply Transition Path Theory (TPT) to drifters originating off the west coast of Africa to the Gulf of Mexico. We find that the most common case of a regular covering by equal longitude-latitude side cells can lead to a large instability in the computed transition times as a function of the number of cells used. We propose a different covering based on a clustering of the trajectory data that is stable against the number of cells in the covering. We also propose a generalization of the standard transition time statistic of TPT that can be used to construct a partition of the domain of interest into weakly dynamically connected regions.

Normal Transaminases in Methamphetamine- and Heroin-Associated Hyperammonemic Encephalopathy.

Detecting undisclosed methamphetamine and heroin abuse is a challenge for life underwriters and medical directors. A common clinical assumption is that if substance abusers experience liver damage, it will be indicated by elevated serum transaminases. The following case suggests that assumption may not be true for heavy substance abusers who consume no or minimal alcohol. This report describes a 44-year-old male with long-term use of inhaled combined methamphetamine and heroin ("speedballs") and minimal alcohol use, whose transaminases remained normal while episodes of acute liver failure and transient hepatic encephalopathy from hyperammonemia were observed. In this case, a fatal motor vehicle accident occurred following the sudden onset of hepatic encephalopathy hours after consuming a "speedball." Normal transaminases may not be proof of a normal healthy liver among methamphetamine and heroin abusers.

Coherent Lagrangian swirls among submesoscale motions.

The emergence of coherent Lagrangian swirls (CLSs) among submesoscale motions in the ocean is illustrated. This is done by applying recent nonlinear dynamics tools for Lagrangian coherence detection on a surface flow realization produced by a data-assimilative submesoscale-permitting ocean general circulation model simulation of the Gulf of Mexico. Both mesoscale and submesoscale CLSs are extracted. These extractions prove the relevance of coherent Lagrangian eddies detected in satellite-altimetry-based geostrophic flow data for the arguably more realistic ageostrophic multiscale flow.

Bioconjugate Supramolecular Pd2+ Metallacages Penetrate the Blood Brain Barrier In Vitro and In Vivo.

The biomedical application of discrete supramolecular metal-based structures, specifically self-assembled metallacages, is still an emergent field of study. Capitalizing on the knowledge gained in recent years on the development of 3-dimensional (3D) metallacages as novel drug delivery systems and theranostic agents, we explore here the possibility to target [Pd2L4]4+ cages (L = 3,5-bis(3-ethynylpyridine)phenyl ligand) to the brain. In detail, a new water-soluble homoleptic cage (CPepH3) tethered to a blood brain barrier (BBB)-translocating peptide was synthesized by a combination of solid-phase peptide synthesis (SPPS) and self-assembly procedures. The cage translocation efficacy was assessed by inductively coupled mass spectrometry (ICP-MS) in a BBB cellular model in vitro. Biodistribution studies of the radiolabeled cage [[99mTcO4]- ⊂ CPepH3] in the CD1 mice model demonstrate its brain penetration properties in vivo. Further DFT studies were conducted to model the structure of the [[99mTcO4]- ⊂ cage] complex. Moreover, the encapsulation capabilities and stability of the cage were investigated using the [ReO4]- anion, the "cold" analogue of [99mTcO4]-, by 1H NMR spectroscopy. Overall, our study constitutes another proof-of-concept of the unique potential of supramolecular coordination complexes for modifying the physiochemical and biodistribution properties of diagnostic species.

In Vivo Pretargeting Based on Cysteine-Selective Antibody Modification with IEDDA Bioorthogonal Handles for Click Chemistry.

Pretargeted imaging has emerged as an effective multistep strategy aiming to improve imaging contrast and reduce patient radiation exposure through decoupling of the radioactivity from the targeting vector. The inverse electron-demand Diels-Alder (IEDDA) reaction between a trans-cyclooctene (TCO)-conjugated antibody and a labeled tetrazine holds great promise for pretargeted imaging applications due to its bioorthogonality, rapid kinetics under mild conditions, and formation of stable products. Herein, we describe the use of functionalized carbonylacrylic reagents for site-specific incorporation of TCO onto a human epidermal growth factor receptor 2 (HER2) antibody (THIOMAB) containing an engineered unpaired cysteine residue, generating homogeneous conjugates. Precise labeling of THIOMAB-TCO with a fluorescent or radiolabeled tetrazine revealed the potential of the TCO-functionalized antibody for imaging the HER2 after pretargeting in a cellular context in a HER2 positive breast cancer cell line. Control studies with MDA-MD-231 cells, which do not express HER2, further confirmed the target specificity of the modified antibody. THIOMAB-TCO was also evaluated in vivo after pretargeting and subsequent administration of an 111In-labeled tetrazine. Biodistribution studies in breast cancer tumor-bearing mice showed a significant activity accumulation on HER2+ tumors, which was 2.6-fold higher than in HER2- tumors. Additionally, biodistribution studies with THIOMAB without the TCO handle also resulted in a decreased uptake of 111In-DOTA-Tz on HER2+ tumors. Altogether, these results clearly indicate the occurrence of the click reaction at the tumor site, i.e., pretargeting of SK-BR-3 HER2-expressing cells with THIOMAB-TCO and reaction through the TCO moiety present in the antibody. The combined advantages of site-selectivity and stability of TCO tagged-antibodies could allow application of biorthogonal chemistry strategies for pretargeting imaging with minimal side-reactions and background.

Transition paths of marine debris and the stability of the garbage patches.

We used transition path theory (TPT) to infer "reactive" pathways of floating marine debris trajectories. The TPT analysis was applied on a pollution-aware time-homogeneous Markov chain model constructed from trajectories produced by satellite-tracked undrogued buoys from the National Oceanic and Atmospheric Administration's Global Drifter Program. The latter involved coping with the openness of the system in physical space, which further required an adaptation of the standard TPT setting. Directly connecting pollution sources along coastlines with garbage patches of varied strengths, the unveiled reactive pollution routes represent alternative targets for ocean cleanup efforts. Among our specific findings we highlight: constraining a highly probable pollution source for the Great Pacific garbage patch; characterizing the weakness of the Indian Ocean gyre as a trap for plastic waste; and unveiling a tendency of the subtropical gyres to export garbage toward the coastlines rather than to other gyres in the event of anomalously intense winds.

Improved Fmoc-solid-phase peptide synthesis of an extracellular loop of CFTR for antibody selection by the phage display technology.

Cystic fibrosis (CF), a life-shortening genetic disease, is caused by mutations in the CF transmembrane conductance regulator (CFTR) gene that codes for the CFTR protein, the major chloride channel expressed at the apical membrane of epithelial cells. The development of an imaging probe capable of non-invasively detect CFTR at the cell surface could be of great advantage for the management of CF. With that purpose, we synthesized the first extracellular loop of CFTR protein (ECL1) through fluorenylmethyloxycarbonyl (Fmoc)-based microwave-assisted solid-phase peptide synthesis (SPPS), according to a reported methodology. However, aspartimide formation, a well-characterized side reaction in Fmoc-SPPS, prompted us to adopt a different side-chain protection strategy for aspartic acid residues present in ECL1 sequence. The peptide was subsequently modified via PEGylation and biotinylation, and cyclized through disulfide bridge formation, mimicking the native loop conformation in CFTR protein. Herein, we report improvements in the synthesis of the first extracellular loop of CFTR, including peptide modifications that can be used to improve antigen presentation in phage display for selection of novel antibodies against plasma membrane CFTR.

Stability of the Malvinas Current.

Deterministic and probabilistic tools from nonlinear dynamics are used to assess enduring near-surface Lagrangian aspects of the Malvinas Current. The deterministic tools are applied to a multiyear record of velocities derived from satellite altimetry data, revealing a resilient cross-stream transport barrier. This is composed of shearless-parabolic Lagrangian coherent structures (LCSs), which, extracted over sliding time windows along the multiyear altimetry-derived velocity record, lie in near-coincidental position. The probabilistic tools are applied on a large collection of historical satellite-tracked drifter trajectories, revealing weakly communicating flow regions as basins of attraction for long-time asymptotic almost-invariant sets on either side of the altimetry-derived barrier. Shearless-parabolic LCSs are detected for the first time from altimetry data, and their significance is supported on satellite-derived ocean color data, which reveal shapes that quite closely resemble the peculiar V shapes, dubbed "chevrons," that have recently confirmed the presence of similar LCSs in the atmosphere of Jupiter. Finally, using available in situ velocity and hydrographic data, sufficient and necessary conditions for nonlinear symmetric stability are found to be satisfied, suggesting a duality between Lagrangian and Eulerian stability for the Malvinas Current.

Markov-chain-inspired search for MH370.

Markov-chain models are constructed for the probabilistic description of the drift of marine debris from Malaysian Airlines flight MH370. En route from Kuala Lumpur to Beijing, MH370 mysteriously disappeared in the southeastern Indian Ocean on 8 March 2014, somewhere along the arc of the 7th ping ring around the Inmarsat-3F1 satellite position when the airplane lost contact. The models are obtained by discretizing the motion of undrogued satellite-tracked surface drifting buoys from the global historical data bank. A spectral analysis, Bayesian estimation, and the computation of most probable paths between the Inmarsat arc and confirmed airplane debris beaching sites are shown to constrain the crash site, near 25°S on the Inmarsat arc.

2014 CRL Build Study of Life Insurance Applicants.

Objective .- Determine the impact of build on insurance applicant mortality accounting for smoking, laboratory test values and blood pressure. Method .- The study consisted of 2,051,370 applicants tested at Clinical Reference Laboratory between 1993 and 2007 with build and cotinine measurements available whose body mass index (BMI) was between 15 and 47. Vital status was determined as of September, 2011 by the Social Security Death Master File. Excluded from the primary study were applicants with HbA1c values ≥6.5%, systolic BP ≥141 mmHg, albumin values ≤3.3 g/dL or total cholesterol values ≤130 mg/dL. Relative mortality was determined by Cox regression analysis for bands of BMI split by age, sex and smoking status (urine cotinine positive). Results .- A majority of applicants had BMI >24 (overweight or obese by WHO criteria). After the exclusions noted above, relative mortality does not increase by >34% unless BMI is <20 (<18 for female non-smokers age 18 to 59) or BMI is >34. BMI values in the range of 22 to 24 and 25 to 29, overall, had similar and the lowest relative risks. For most nonsmokers, risk was lowest in the lower of these two BMI bands but for smokers (and non-smoking males age 60 to 89) risk was lowest in the higher BMI band. Additional analysis showed limited reduction in relative risk by accounting for all laboratory test values as well as continuing the exclusions. Eliminating the exclusions resulted in only a modest increase in relative risk because the mortality rate of the reference band increased as well. Conclusion .- After excluding elevated HbA1c and blood pressure (associated with high BMI) and low albumin and cholesterol (associated with low BMI) which are usually evaluated separately, mortality varies by a limited degree for BMI 20 to 34. Accounting for the mortality impact of other test values, in addition to the exclusions noted, reduced mortality associated with high BMI to a limited extent, but had little impact on mortality associated with low BMI.

Molecular characterization of infectious bursal disease virus (IBDV) isolated in Argentina indicates a regional lineage.

In Argentina, classical vaccines are used to control infectious bursal disease virus (IBDV); however, outbreaks of IBDV are frequently observed. This could be due to failures in the vaccination programs or to the emergence of new strains, which would be able to break through the protection given by vaccines. Hence, genetic characterization of the viruses responsible for the outbreaks that occurred in recent years is crucial for the evaluation of the control programs and the understanding of the epidemiology and evolution of IBDV. In this study, we characterized 51 field samples collected in Argentina (previously identified as IBDV positive) through the analysis of previously identified apomorphic sequences. Phylogenetic analysis of regVP2 showed that 42 samples formed a unique cluster (Argentinean lineage), seven samples were typical classical strains (one of them was a vaccine strain), and two belonged to the very virulent lineage (vvIBDV). Interestingly, when the analysis was performed on the regVP1 sequences, the field samples segregated similarly to regVP2; thus, we observed no evidence of a reassortment event in the Argentinean samples. Amino acid sequence analysis of regVP2 showed a particular pattern of residues in the Argentinean lineage, particularly the presence of T272, P289 and F296, which had not been reported before as signature sequences for any IBDV phenotype. Notably, the residue S254, characteristic of the antigenic variant, was not present in any of the Argentinean samples.

Isostructural Re(I)/(99m)Tc(I) tricarbonyl complexes for cancer theranostics.

Merging classical organic anticancer drugs with metal-based compounds in one single molecule offers the possibility of exploring new approaches for cancer theranostics, i.e. the combination of diagnostic and therapeutic modalities. For this purpose, we have synthesized and biologically evaluated a series of Re(I)/(99m)Tc(I) tricarbonyl complexes (Re1­Re4 and Tc1­Tc4, respectively) stabilized by a cysteamine-based (N,S,O) chelator and containing 2-(4'-aminophenyl)benzothiazole pharmacophores. With the exception of Re1, all the Re complexes have shown a moderate cytotoxicity in MCF7 and PC3 cancer cells (IC50 values in the 15.9­32.1 µM range after 72 h of incubation). The cytotoxic activity of the Re complexes is well correlated with cellular uptake that was quantified using the isostructural (99m)Tc congeners. There is an augmented cytotoxic effect for Re3 and Re4 (versusRe1 and Re2), and the highest cellular uptake for Tc3 and Tc4, which display a long ether-containing linker to couple the pharmacophore to the (N,S,O)-chelator framework. Moreover, fluorescence microscopy clearly confirmed the cytosolic accumulation of the most cytotoxic compound (Re3). Biodistribution studies of Tc1­Tc4 in mice confirmed that these moderately lipophilic complexes (logDo/w = 1.95­2.32) have a favorable bioavailability. Tc3 and Tc4 presented a faster excretion, as they undergo metabolic transformations, in contrast to complexes Tc1 and Tc2. In summary, our results show that benzothiazole-containing Re(I)/(99m)Tc(I) tricarbonyl complexes stabilized by cysteamine-based (N,S,O)-chelators have potential to be further applied in the design of new tools for cancer theranostics.

Neuronal deletion of GSK3ß increases microtubule speed in the growth cone and enhances axon regeneration via CRMP-2 and independently of MAP1B and CLASP2.

BACKGROUND: In the adult central nervous system, axonal regeneration is abortive. Regulators of microtubule dynamics have emerged as attractive targets to promote axonal growth following injury as microtubule organization is pivotal for growth cone formation. In this study, we used conditioned neurons with high regenerative capacity to further dissect cytoskeletal mechanisms that might be involved in the gain of intrinsic axon growth capacity. RESULTS: Following a phospho-site broad signaling pathway screen, we found that in conditioned neurons with high regenerative capacity, decreased glycogen synthase kinase 3ß (GSK3ß) activity and increased microtubule growth speed in the growth cone were present. To investigate the importance of GSK3ß regulation during axonal regeneration in vivo, we used three genetic mouse models with high, intermediate or no GSK3ß activity in neurons. Following spinal cord injury, reduced GSK3ß levels or complete neuronal deletion of GSK3ß led to increased growth cone microtubule growth speed and promoted axon regeneration. While several microtubule-interacting proteins are GSK3ß substrates, phospho-mimetic collapsin response mediator protein 2 (T/D-CRMP-2) was sufficient to decrease microtubule growth speed and neurite outgrowth of conditioned neurons and of GSK3ß-depleted neurons, prevailing over the effect of decreased levels of phosphorylated microtubule-associated protein 1B (MAP1B) and through a mechanism unrelated to decreased levels of phosphorylated cytoplasmic linker associated protein 2 (CLASP2). In addition, phospho-resistant T/A-CRMP-2 counteracted the inhibitory myelin effect on neurite growth, further supporting the GSK3ß-CRMP-2 relevance during axon regeneration. CONCLUSIONS: Our work shows that increased microtubule growth speed in the growth cone is present in conditions of increased axonal growth, and is achieved following inactivation of the GSK3ß-CRMP-2 pathway, enhancing axon regeneration through the glial scar. In this context, our results support that a precise control of microtubule dynamics, specifically in the growth cone, is required to optimize axon regrowth.

2014 CRL Blood Pressure Study of Life Insurance Applicants.

Objective .- Define the relative mortality risk by systolic (SBP) and diastolic blood pressure (DBP) in a relatively healthy cohort split by age and sex with adjustment for smoking status, other findings and admitted heart disease history. Method .- Blood pressure (BP in mm Hg), build, laboratory studies and limited medical history are collected when people apply for individual life insurance. Information on 2,472,706 applicants tested by Clinical Reference Laboratory from 1993 to 2007 was utilized with follow-up for vital status using the September 2011 Social Security Death Master File identifying 31,033 deaths. Data was analyzed by SBP and DBP split by age and sex accounting for smoking and for BMI, urine protein/creatinine ratio and history of heart disease in a Cox multivariate survival analysis. Separate analysis by admitted hypertension history was also conducted. Results are presented by SBP and DBP for 4 age-sex groups with and without added covariates beyond age and smoking status. Results .- Relative mortality progressively increased by SBP level from the 90 to 119 band (down to 80 in younger women) upward with little additional impact by DBP. Addition of covariates beyond age and smoking resulted in a 5% to 10% reduction in relative risk. Although high DBP had limited impact, a pulse pressure/SBP ratio >½ identified 1% of applicants at high mortality risk, with little difference in risk for ratios ≤½. Hypertension history with current BP control was associated with a 10% to 25% increase in relative mortality risk as compared to those with similar BP but no such history. Conclusion .- Increasing SBP is closely associated with increasing relative mortality, starting from the lowest SBP. Increasing DBP has little additional impact, but a pulse pressure/SBP ratio >½ is a potent marker of increased risk as well. Accounting for build and other laboratory findings reduces risk modestly. A history of hypertension with current control increases risk.

Changing the "Normal Range" for Blood Pressure from 140/90 to 130/Any Improves Risk Assessment.

Objective .- Redefine the "normal" reference range for blood pressure from <140/90 to one that more effectively identifies individuals with increased mortality risk. Method .- Data from the recently published 2014 CRL blood pressure study was used. It includes 2,472,706 life insurance applicants tested by Clinical Reference Laboratory from 1993 to 2007 with follow-up for vital status using the September 2011 Social Security Death Master File. Various upper limits of blood pressure (BP in mm Hg) were evaluated to determine if any was superior to the current, commonly used limit of 140/90 in identifying individuals with increased mortality risk. Results .- An alternative reference range using a systolic BP (SBP) <130 with any diastolic BP (DBP) included 84% of life insurance applicants. It had a lower mortality rate and narrower range of relative risk than <140/90, including 89% as many applicants but only 68% as many deaths. This pattern of lives and deaths was consistent across age and sex. Conclusion .- Switching to a "normal" reference range of SBP <130 offers superior risk assessment relative to using BP <140/90 while still including a sufficient percentage of the population.

Hemoglobin Screening Independently Predicts All-Cause Mortality.

Objective .- Determine if the addition of hemoglobin testing improves risk prediction for life insurance applicants. Method .- Hemoglobin results for insurance applicants tested from 1993 to 2007, with vital status determined by Social Security Death Master File follow-up in 2011, were analyzed by age and sex with and without accounting for the contribution of other test results. Results .- Hemoglobin values ≤12.0 g/dL (and possibly ≤13.0 g/dL) in females age 50+ (but not age <50) and hemoglobin values ≤13.0 g/dL in all males are associated with progressively increasing mortality risk independent of the contribution of other test values. Increased risk is also noted for hemoglobin values >15.0 g/dL (and possibly >14.0 g/dL) for all females and for hemoglobin values >16.0 g/dL for males. Conclusion .- Hemoglobin testing can add additional independent risk assessment to that obtained from other laboratory testing, BP and build in this relatively healthy insurance applicant population. Multiple studies support this finding at older ages, but data (and the prevalence of diseases impacting hemoglobin levels) are limited at younger ages.

Membrane lipid profile alterations are associated with the metabolic adaptation of the Caco-2 cells to aglycemic nutritional condition.

Cancer cells can adapt their metabolic activity under nutritional hostile conditions in order to ensure both bioenergetics and biosynthetic requirements to survive. In this study, the effect of glucose deprivation on Caco-2 cells bioenergetics activity and putative relationship with membrane lipid changes were investigated. Glucose deprivation induces a metabolic remodeling characterized at mitochondrial level by an increase of oxygen consumption, arising from an improvement of complex II and complex IV activities and an inhibition of complex I activity. This effect is accompanied by changes in cellular membrane phospholipid profile. Caco-2 cells grown under glucose deprivation show higher phosphatidylethanolamine content and decreased phosphatidic acid content. Considering fatty acid profile of all cell phospholipids, glucose deprivation induces a decrease of monounsaturated fatty acid (MUFA) and n-3 polyunsaturated fatty acids (PUFA) simultaneously with an increase of n-6 PUFA, with consequent drop of n-3/n-6 ratio. Additionally, glucose deprivation affects significantly the fatty acid profile of all individual phospholipid classes, reflected by an increase of peroxidability index in zwitterionic phospholipids and a decrease in all anionic phospholipids, including mitochondrial cardiolipin. These data indicate that Caco-2 cells metabolic remodeling induced by glucose deprivation actively involves membrane lipid changes associated with a specific bioenergetics profile which ensure cell survival.